Related Experiment Video
Updated: Jul 9, 2026
18:07
Analyzing Cellular Internalization of Nanoparticles and Bacteria by Multi-spectral Imaging Flow Cytometry
Published on: June 8, 2012
Method for nanoparticles uptake evaluation based on double labeled fluorescent cells scanned in enhanced darkfield
Mona Mihailescu1,2, Luminita C Miclea3,4, Ana M Pleava5,6
1Holographic Imaging and Processing Laboratory, Physics Department, Politehnica University Bucharest, 313 Splaiul Independentei, Bucharest, 060042, Romania.
Biomedical Optics Express
|June 21, 2023
Summary
This study introduces a new method for tracking nanoparticle (NP) uptake in cells using advanced imaging and computational analysis. The findings reveal NP saturation limits and decreased cell viability at high concentrations.
Area of Science:
- Nanotechnology
- Cell Biology
- Biophysics
Background:
- Accurate quantification of intracellular nanoparticle (NP) uptake is crucial for understanding NP-cell interactions and their toxicological effects.
- Existing imaging techniques often lack the resolution or computational power to precisely track NPs within distinct cellular compartments.
Purpose of the Study:
- To develop and validate an integrated imaging and computational method for quantitative analysis of nanoparticle traffic within cells.
- To assess NP uptake efficiency, distribution, and impact on cell viability across different cellular regions.
Main Methods:
- Utilized an enhanced dark field CytoViva optical system combined with 3D reconstructions of fluorescently labeled cells and hyperspectral imaging.
- Developed MATLAB scripts for image processing, partitioning cell volumes into nucleus, cytoplasm, and membrane-associated shells, and counting NPs in each region.
- Computed parameters including regional NP densities, flow densities, relative accumulation indices, and uptake ratios.
Main Results:
- The method successfully quantified NP localization and traffic within defined cellular compartments, aligning with biochemical analyses.
- Demonstrated a saturation limit for intracellular NP density at high extracellular NP concentrations.
- Observed higher NP densities near plasma membranes and a decrease in cell viability correlated with increasing extracellular NP concentration.
Conclusions:
- The developed method provides a robust platform for evaluating intracellular nanoparticle dynamics and their biological consequences.
- High extracellular NP concentrations can lead to cellular saturation, reduced viability, and altered cell morphology.
- The findings highlight the importance of considering NP concentration-dependent effects on cellular uptake and viability.
